论文部分内容阅读
目的通过应用抗氧化剂葡萄籽初步探讨其能否通过抑制肌细胞凋亡来延缓失神经支配后骨骼肌的萎缩现象。方法选择8周龄Vistar大鼠40只,随机分为A组(对照组)、B组(去神经14d组)、C组(去神经28d组)、D组(去神经14d干预组)、E组(去神经28d干预组)等5组,A组做假手术,B、C、D、E组制作坐骨神经切断后失神经支配的腓肠肌模型。术后D、E组用葡萄籽200mg.kg-1.d-1空腹灌胃,A、B、C组用等量生理盐水灌胃。术后分别于14d、28d处死大鼠,用流式细胞仪检测腓肠肌细胞活性氧(ROS)的含量,激光共聚焦显微镜检测腓肠肌细胞线粒体通透性转化孔(MPTP)的开放情况,脱氧核糖核苷酸转移酶介导的缺口末端标记(Tunel)法检测腓肠肌细胞凋亡,肌湿重之比评价骨骼肌萎缩情况。结果大鼠失神经支配后,B、C、D、E组肌细胞中的ROS、MPTP及凋亡率与正常对照组比较,表达随去神经支配时间的延长(<28d)而持续增加,且差异具有统计学意义(P<0.05),D、E组肌细胞ROS、MPTP及肌凋亡率分别与B、C组比较,表达有所减少,差异具有统计学意义(P<0.05);B、C组肌湿重之比较A组显著减小(P<0.05),D、E组肌湿重比分别与B、C组相比,有所升高,且差异具有统计学意义(P<0.05)。结论 ROS为调控失神经支配后骨骼肌萎缩的重要分子,葡萄籽可以通过减少ROS的含量进而抑制肌细胞凋亡并最终延缓骨骼肌萎缩。
OBJECTIVE: To investigate whether it can delay the atrophy of skeletal muscle after denervation by inhibiting the apoptosis of muscle cells through the application of antioxidant grape seed. Methods Forty eight-week-old Vistar rats were randomly divided into three groups: control group A, control group, group B (denervation 14d group), group C (denervation 28d group), group D Group (denervation 28d intervention group) and other 5 groups, A group of sham operation, B, C, D, E group produced sciatic nerve after denervation dominated gastrocnemius model. Postoperative D, E group with grape seed 200mg.kg-1.d-1 fasting gavage, A, B, C group with the same amount of normal saline. The rats were sacrificed on the 14th day and the 28th day respectively. The content of reactive oxygen species (ROS) in gastrocnemius muscle was detected by flow cytometry. The opening of mitochondrial permeability transition pore (MPTP) in gastrocnemius muscle was detected by confocal microscopy. TUNEL-mediated nick end labeling (Tunel) assay gastrocnemius muscle apoptosis, muscle wet weight ratio evaluation of skeletal muscle atrophy. Results After denervation, the ROS, MPTP and apoptosis rates of myocytes in groups B, C, D and E were increased continuously (<28d) compared with the control group (P <0.05). The ROS, MPTP and myocyte apoptosis rate of myocytes in D and E groups were decreased compared with those in B and C groups, the difference was statistically significant (P <0.05); B , The muscle wet weight in group C was significantly lower than that in group A (P <0.05), and the muscle wet weights in groups D and E were increased compared with those in groups B and C, respectively (P < 0.05). Conclusion ROS is an important molecule in regulating skeletal muscle atrophy after denervation. Grape seed can inhibit myocyte apoptosis and decrease skeletal muscle atrophy by reducing ROS content.